Disease Outcome and Brain Metabolomics of Cyclophilin-D Knockout Mice in Sepsis

Sepsis-associated encephalopathy (SAE) is a diffuse brain dysfunction resulting from a systemic inflammatory response to infection, but the mechanism remains unclear. The mitochondrial permeability transition pore (MPTP) could play a central role in the neuronal dysfunction, induction of apoptosis, and cell death in SAE. The mitochondrial isomerase cyclophilin D (CypD) is known to control the sensitivity of MPTP induction. We, therefore, established a cecal ligation and puncture (CLP) model, which is the gold standard in sepsis research, using CypD knockout (CypD KO) mice, and analyzed the disease phenotype and the possible molecular mechanism of SAE through metabolomic analyses of brain tissue. A comparison of adult, male wild-type, and CypD KO mice demonstrated statistically significant differences in body temperature, mortality, and histological changes. In the metabolomic analysis, the main finding was the maintenance of reduced glutathione (GSH) levels and the reduced glutathione/oxidized glutathione (GSH/GSSG) ratio in the KO animals following CLP. In conclusion, we demonstrate that CypD is implicated in the pathogenesis of SAE, possibly related to the inhibition of MPTP induction and, as a consequence, the decreased production of ROS and other free radicals, thereby protecting mitochondrial and cellular function.

Effect of styrene oxide and diethyl maleate on expression of cytochrome P450 family 1 and glutathione store in mouse liver

Purpose: To determine the effect of the glutathione (GSH) suppressors styrene oxide (SO) and diethyl maleate (DEM) on the hepatic expression of cytochrome P450 family 1 (Cyp1) isoforms that are related to carcinogenesis including Cyp1a1, Cyp1a2, and Cyp1b1. Methods: Seven-week-old ICR mice were intraperitoneally injected with SO (150 and 300 mg/kg/day), DEM (175 and 350 mg/kg/day), or N-acetylcysteine (NAC; 300 and 600 mg/kg/day) for 7, 14, or 28 days. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, hepatic Cyp1 expression, total glutathione, reduced glutathione (GSH), and oxidized glutathione (GSSG) were determined. Results: ALT and AST levels were markedly increased by SO and DEM while GSH/GSSG ratio was decreased by SO in all treatments (p < 0.05), while high dose (350 mg/kg/day) DEM significantly suppressed GSH/GSSG ratio at 28 days (p < 0.05). The expressions of Cyp1a1, Cyp1a2, and Cyp1b1 were induced by SO and DEM, corresponding with induction of ethoxy/methoxy-resorufin O- dealkylase activities. Conclusion: The Cyp1 family metabolizes procarcinogens to carcinogenic DNA adducts; exposure to the industrial solvents, SO and DEM, raises the risk of carcinogenesis via GSH depletion coupled with Cyp1 induction.

Effect of antioxidants and growth regulators on shoot organogenesis in the apical meristem culture of Fragaria × ananassa (Duchesne ex Weston) Duchesne ex Rozier

The initiation of strawberries into in vitro culture is known to be complicated by the inhibition of organogenesis by phenolic oxidation products. An important role in this process is given to the selection of growth regulators that increase meristematic cell activity and shoot proliferation at the stage of organogenesis induction. The present study aims to obtain a viable apical meristem culture of garden strawberry and to study the effect of different antioxidants (reduced glutathione (RG); a new preparation, i.e., a mechanical composite (MC) on the basis of biogenic silicon and green tea catechins and plant growth regulators (6-benzylaminopurine; thidiazuron) on the initiation of axillary shoot formation in strawberry meristem culture. Terminal buds containing an apical meristem and two leaf primordia isolated from the stolons of two garden strawberry cultivars (Sunny Meadow and Festival Chamomile) were used as primary explants for the initiation of strawberries into in vitro culture. It was found for the first time that the MC exhibits higher antioxidant activity as compared to reduced glutathione, reduces darkening of initial explants, as well as enhancing regeneration up to 13.0% at p ≤ 0.05. Furthermore, the best effect on the formation of microshoots per explant is observed toward the end of material introduction into in vitro culture when combining the MC with growth regulators in the culture medium. Here, the effect of strawberry cultivar on explant regeneration and the number of microshoots per explant are insignificant. It is concluded that the procedure for using the MC as an effective antioxidant during material initiation into the culture can be applied to the large-scale in vitro propagation of garden strawberries. Moreover, the technology for obtaining the MC from plant waste is environmentally friendly, which is a significant advantage for its use in in vitro technologies.

Toxicity of The Microcystin-Producing Cyanobacterium Microcystis Aeruginosa To Litopenaeus Vannamei

Microcystis aeruginosa is a common kind of harmful bloom algae, which was also frequently found as a dominant microalgae specie in shrimp breeding ponds. And it was found that blooms always induced massive death of shrimp, but the toxic effects of M. aeruginosa on Litopenaeus vannamei are still not completely understood. In this paper, the toxicity of M. aeruginosa cells to L. vannamei was examined, and the toxic components in the cells were analyzed through high-pressure liquid chromatography (HLPC). In addition, the immune response of shrimp to the microalgal extract was assessed by measuring the activity of immune-related enzymes, as well as the transcription of the relevant genes. Overall, both M. aeruginosa cells and the algal extract resulted in a 100% mortality rate in shrimp, whereas the cell-free culture medium was ineffective. And HPLC analysis results revealed the presence of microcystin-LR (MC-LR) at a concentration of 190.40 mg/kg of cells. In addition, the activity and gene transcription of two immune related enzymes, SOD and LZM, were both significantly reduced in shrimp hepatopancreas (p<0.05) after injection with cell extract. However, reduced glutathione (GSH) content was slightly increased, but the ratio of GSH to GSSG was down, and the transcription of gst gene function as detoxification, was significantly downregulated (p<0.05). The results demonstrated that M. aeruginosa cell extract was highly toxic to L. vannamei, and exerted a negative effect on shrimp immunity including reduction of antioxidant capacity, antibacterial activity and detoxification activity, due to microcystin-LR.

Self-Healing Behavior of Polymer/Protein Hybrid Hydrogels

The paper presents the viscoelastic properties of new hybrid hydrogels containing poly(vinyl alcohol) (PVA), hydroxypropylcellulose (HPC), bovine serum albumin (BSA) and reduced glutathione (GSH). After heating the mixture at 55 °C, in the presence of GSH, a weak network is formed due to partial BSA unfolding. By applying three successive freezing/thawing cycles, a stable porous network structure with elastic properties is designed, as evidenced by SEM and rheology. The hydrogels exhibit self-healing properties when the samples are cut into two pieces; the intermolecular interactions are reestablished in time and therefore the fragments repair themselves. The effects of the BSA content, loaded deformation and temperature on the self-healing ability of hydrogels are presented and discussed through rheological data. Due to their versatile viscoelastic behavior, the properties of PVA/HPC/BSA hydrogels can be tuned during their preparation in order to achieve suitable biomaterials for targeted applications.

Evaluation of the antitumor and antioxidant effects of jatobá (Hymenaea courbaril) extracts / Avaliação do efeito antitumoral e antioxidante de extratos do jatobá (Hymenaea courbaril)

Ethnobotanical surveys have revealed the use of jatobá for the treatment of several diseases. This study determined the effect of plant extracts on the development of Ehrlich carcinoma. Male Swiss mice (n=6) were subcutaneously inoculated with 106 tumor cells and intragastrically administered ethanol (2 mg·mL-1, 5 mg·mL-1, or 10 mg·mL-1) or aqueous extracts of jatobá seed or bark for 90 days. Tumor development did not significantly differ between the groups studied; however, animals treated with the aqueous extract of the seed (2.205 mg·mL-1) had a reduction in tumor size compared to those treated with the aqueous extract of the bark (1.7 mg·mL-1). The treatment was not found to influence the survival of the animals studied. A new group of animals (n=7), with or without the tumor, received the aqueous extract of jatobá seed for 7, 14, and 30 days to evaluate oxidative stress. The extract reduced the thiobarbituric acid reactive substances levels at 7 days in the liver and kidneys, and 14 days in brain and renal tissue. Protein carbonylation levels were also reduced at 7 days in the liver and brain tissue and 14 days in the liver. The reduced glutathione levels diminished in animals treated for 7 and 14 days. We conclude that treatment with the aqueous extract of the jatobá seed presents promising activity in the reduction of oxidative stress.

Antioxidant status in children and adolescents with COVID-19

Background. The COVID-19 pandemic has raised the importance of this problem to the first stage and has affected healthcare system around the world. Despite the more favorable COVID-19 course, the child population should be at focus of special attention, due to the active participation in its distribution. The course of COVID-19 includes a cascade of pathological processes accompanied by the generation of reactive oxygen species, which can have extremely negative consequences for the developing organism. The research of these processes in children is vital and will improve the effectiveness of preventive and therapeutic measures. The aim: to analyze changes in enzymatic and non-enzymatic links in the antioxidant defense in children and adolescents with diagnosed COVID-19 infection.Materials and methods. 17 children and adolescents (average age – 12.35 ± 4.01 years) were examined, including 8 boys (47 %) and 9 girls (53 %) with COVID-19 infection. The control group of children and adolescents (practically healthy) according to the «copy-pair» principle was selected. We used spectrophotometric methods.Results. In the group of children and adolescents with diagnosed COVID-19 infection, there were lower levels of total antioxidant activity (p < 0.0001), superoxide dismutase activity (p < 0.0001), content of reduced glutathione (p = 0.048) and retinol (p = 0.015), increase in glutathione reductase activity (p = 0.015) relative to the control.Conclusion. The obtained data indicate the insufficiency of antioxidant system components number in children and adolescents with diagnosed COVID-19 infection and indicate the advisability of antioxidant therapy using to stabilize these indicators.

Hepatoprotective Activity of Helicteres isora Ethanol Extract Against Paracetamol-Induced Liver Injury in Mice

Helicteres isora L. is a medicinal plant which is used in several diseases, such as snake-bite, dog-bite, diarrhoea and constipation in a new born baby, gastrointestinal disorders, diabetes, cancer, and infections. This plant has also been used in the management of liver damage through traditional medicine. However, the hepatoprotective activity of H. isora L. ethanolic extract has not been reported so much. The present work was carried to investigate the hepatoprotective effect of H. isora L. against paracetamol-induced liver injury in Swiss mice. Paracetamol (PCM) is widely used as an analgesic and antipyretic drug which at high dose can lead to undesirable side effects, such as hepatotoxicity. Paracetamol induce hepatotoxicity was evaluated by an increase (P<0.05) in AST and ALT serum activity. Paracetamol hepatotoxicity was also manifested by an increase in (P<0.05) lipid peroxidation and depletion of reduced glutathione (GSH) in liver tissue. Ethanol extract of H. isora L. (250, 500 and 1000 mg/kg bw/day) significantly restored the PCM-induced alterations in the biochemical activities of blood and liver tissues. The hepatoprotective effect of H. isora L. was also confirmed by the histopathological examination of liver tissue. Histopathological examination of liver sections in mice administered with 1000 mg/kg bw/day doses of the extract were perfectly protected almost similar to those of untreated mice. The results indicated the hepatoprotective nature of studied plants extract against paracetamol induced toxicity. Our study scientifically validates the folkloric claim as well as traditional uses of H. isora L. as hepatoactive medicine. The results of this study suggests a new direction in the treatment of liver disease in future.

Protective Effect of Gallic Acid Against Thioacetamide Induced Metabolic Dysfunction of Lipids and Hepatic and Renal Toxicity

BackgroundGallic acid (GA) has a potential antioxidant bio-activity and inhibits diet-induced hypertriglyceridemia with reducing the size of adipocytes. GA also was found to increase the uptake of glucose by cell.MethodsThe present research studied the influence of gallic acid (GA) (100mg, 200 mg/Kg orally) on the liver and kidney injuries motivated by thioacetamide (TAA; 100 mg/Kg IP). The treatment of TAA was carried out three times weekly for eight weeks, while gallic acid was given daily. ResultsGA relieved the decrease of hepatic or renal reduced glutathione (GSH) or increase of malondialdehyde (MDA, an indicator for lipid peroxidation) induced by TAA. TAA treatment led to a significant increase in plasma inflammatory markers (TNF-α, CRP), liver enzymes (Gamma-glutamyltransferase (GGT), Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), alkaline phosphatase (ALP) and kidney function parameters (creatinine, urea, uric acid). However, these parameters were reduced after treatment with GA. Moreover, GA reduced the significant decline in plasma protein induced by TAA. In addition, the hepatic fibrosis or histopathological changes of the liver and kidney were lowered by GA. ConclusionOur results suggested that GA may attenuate TAA induced liver and kidney toxicity via suppression of oxidative stress.

Glutathione-dependent system in the blood of gastric cancer patients with various tumor histotypes and prevalence of the disease

Purpose of the study. Exploring the erythrocyte glutathione system functioning in patients with gastric cancer in a comparative aspect, depending on tumor histotype and disease prevalence.Patients and methods. The study included 89 patients with gastric cancer, divided into 6 groups depending on the histotype of the tumor. Separately, the results of the study were analyzed in patients with T4 status according to TNM and in patients at stage 4. The content of reduced glutathione and the activity of glutathione-dependent enzymes in the erythrocytes of the patients' blood were studied by conventional spectrophotometric methods. Statistical processing of the results was carried out using the Statistika 6.0 software package according to the Student's t-test and the nonparametric Wilcoxon-Mann-Whitney test for two independent samples.Results. An increase in the content of glutathione was revealed in patients with gastric cancer compared with the group without oncopathology. The maximum increase was observed in patients with low-grade adenocarcinoma - by 42.5 %, while in signet ring cell carcinoma (SRCC) there was only a tendency to increase by 17.8 %. Glutathione reductase activity was decreased in adenocarcinoma by 23.4-26.2 % and did not change in SRCC. The activity of the antioxidant enzymes glutathione peroxidase and glutathione transferase was increased in all groups, and especially in SRCC - by 76 % and 23-29 %, respectively. In patients with the T4 status and at stage IV of the process, a lower activity of the studied glutathione-dependent enzymes was revealed in comparison with all other groups of patients.Conclusions. The data obtained indicate a greater functional potential of the glutathione system in SRCC. A significant increase in glutathione transferase activity at a sufficiently high level of reduced glutathione can contribute to the development of treatment resistance in SRCC patients.